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Bordetella Pertussis
Bordetella Pertussis
1. Microscopy:
▪ Though the disease is mainly in the lower respiratory tract, the organism can be
recovered readily from the nasopharynx.
▪ ‘Cough plates’ and postnasal swabs are unsatisfactory because of overgrowth by
commensal bacteria.
▪ The optimal diagnostic specimen is a naso-pharyngeal aspirate.
▪ i. The Cough Plate Method
▪ Here a culture plate is held about 10-15 cm in front of the patient’s mouth
during about of spontaneous or induced coughing so that droplets of
respiratory exudates impinge directly on the medium.
▪ This has the advantage that specimen is directly inoculated at the bedside.
▪ ii. The Postnasal (Peroral) Swab
▪ Secretions from the posterior pharyngeal wall are collected with a cotton
swab on a bent wire passed through the mouth.
▪ Salivary contamination should be avoided.
▪ A West’s postnasal swab may be conveniently employed.
▪ Cotton swabs should not be used because they contain fatty acids that are
toxic to B. pertussis so it is preferable to use dacron or calcium alginate
swabs for specimen collection.
▪ iii. The Pernasal Swab
▪ A sterile swab on a flexible wire is passed gently along the floor of the nose
until it meets resistance.
▪ The swab, which will collect mucopus, is withdrawn and either plated
immediately on charcoal blood agar, or placed in transport medium.
▪ The use of transport medium reduces the isolation rate.
▪ A single swab may yield a negative culture, but isolation rates of up to 80
percent may be achieved by taking specimens on several successive days.
▪ The pernasal swab has generally replaced the cough plates or post- nasal
swabs that were used in the past.
3. Culture:
▪ The swab is inoculated immediately on charcoalhorse blood agar and Bordet-Gengou
medium both with and without methicillin or cephalexin and incubated for at least
seven days before being discarded as negative.
▪ The specimen may be transported in Regan-Lowe semi-solid medium if delay in
transport is unavoidable.
▪ Plates are incubated in high humidity at 35-36°C and colonies appear in 48-72 hours.
▪ Typical ‘bisected pearl’ colonies appearing after 3-5 days must be investigated
further.
4. Identification:
▪ Bordetella antigens may be detected in serum and urine in tests with specific
antiserum.
▪ Alternatively, bacteria in nasopharyngeal secretions are labelled with fluorescein-
conjugated antiserum and examined by ultraviolet microscopy.
▪ This method has the theoretical advantage, compared with culture, of detecting dead
bordetellae.
▪ Polymerase chain reaction (PCR) is used for the detection of bordetella DNA in
nasopharyngeal specimens, by the use of various primers with a sensitivity of 80
percent to 100 percent.
7. Serology:
• BORDETELLA PERTUSSIS
• DPT VACCINE
• WHOOPING COUGH